From owner-7tms_r@net.bio.net Wed Nov 01 22:00:00 1995 Path: biosci!bath.ac.uk!bssggl From: bssggl@bath.ac.uk (G G Lunt) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe and info Date: 2 Nov 1995 01:14:50 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 4 Sender: daemon@net.bio.net Distribution: world Message-ID: Please can I have details of joining 7tms. Also - does anyone out there have good models of the dopamine receptor family binding sites into which we might dock some interesting ligands?? George Lunt & Marcela Aliste, University of Bath, UK From owner-7tms_r@net.bio.net Thu Nov 02 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!news00.sunet.se!sunic!news99.sunet.se!news.funet.fi!news.abo.fi!usenet From: Tuomo Glumoff Newsgroups: bionet.molbio.proteins.7tms_r Subject: Sucrose monolaurate Date: 3 Nov 1995 12:29:27 GMT Organization: Abo Akademi University Lines: 6 Message-ID: <47d1v7$pu6@josie.abo.fi> NNTP-Posting-Host: tuomo.btk.utu.fi Does anybody recommend Sucrose monolaurate for solubilization of integral membrane proteins? References? Thanks! Tuomo Glumoff From owner-7tms_r@net.bio.net Wed Nov 08 22:00:00 1995 Path: biosci!rutgers!uwm.edu!chi-news.cic.net!simtel!swidir.switch.ch!scsing.switch.ch!news.belwue.de!news.uni-ulm.de!rz.uni-karlsruhe.de!news.uni-stuttgart.de!uni-regensburg.de!lrz-muenchen.de!news From: diabetes@lrz.uni-muenchen.de (Wolfgang Schechinger) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Q What stands the 7tms_r for? Date: Thu, 09 Nov 1995 21:01:29 GMT Organization: Institute for Diabetes Research Lines: 15 Distribution: world Message-ID: <47tmrt$nr2@sparcserver.lrz-muenchen.de> NNTP-Posting-Host: u7k0201.ppp.lrz-muenchen.de X-Newsreader: Forte Free Agent 1.0.82 +++++++++++++++++++++++++++++++++++++++++++ Wolfgang Schechinger Chemist, PhD Student Institute for Diabetes Reserch Koelner Platz 1 80804 Munich Germany Phone +49 (89) 30 79 31 24 Fax +49 (89) 30 81 733 email u7k0201@sunmail.lrz-muenchen.de (Standard disclaimer applies) +++++++++++++++++++++++++++++++++++++++++++ From owner-7tms_r@net.bio.net Wed Nov 08 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!chi-news.cic.net!simtel!swidir.switch.ch!scsing.switch.ch!news.belwue.de!news.uni-ulm.de!rz.uni-karlsruhe.de!news.uni-stuttgart.de!uni-regensburg.de!lrz-muenchen.de!faui0n.informatik.uni-erlangen.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!krasel From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Q What stands the 7tms_r for? Date: 9 Nov 1995 22:18:46 GMT Organization: Dept. of Pharmacology, U Wuerzburg Lines: 13 Distribution: world Message-ID: <47tuo6$ric@winx03.informatik.uni-wuerzburg.de> References: <47tmrt$nr2@sparcserver.lrz-muenchen.de> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] Wolfgang Schechinger (diabetes@lrz.uni-muenchen.de) wrote: in the subject: > What stands the 7tms_r for? Seven-transmembrane-segment receptors (or something very similar). --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Sat Nov 11 22:00:00 1995 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: IMPORTANT: BIOSCI miniFAQ Date: 12 Nov 1995 02:01:23 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 196 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511121000.CAA05153@net.bio.net> NNTP-Posting-Host: net.bio.net This is a new "miniFAQ" designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. Contents: -------- 1) What to do about "spams," i.e., junk mail, ads, etc. 2) Examples of subscribing and unsubscribing to the mailing lists. 3) How to access BIOSCI/bionet newsgroup archives. 4) The BIOSCI user address and research interest directory. 1) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups) and mailing lists. The same postings are distributed on both media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the newsgroups from about 95% of the spams that are being sent to date. This means that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings. Unfortunately there are easy ways for determined spammers to override the moderation mechanism. We are working on new systems to provide access to our newsgroups over the WWW. These should be available soon, probably November 1995, and will allow you to use your Web browser to look at the news postings. While this will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. 2) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. 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For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 3) How to access BIOSCI/bionet newsgroup archives. -------------------------------------------------- Back postings of all BIOSCI/bionet newsgroups can be found on the World Wide Web at URL http://www.bio.net/. There are several searchable newsgroup indices at this site. E-mail users can search the BIOSCI archives by using our waismail e-mail server. For instructions send the message help to waismail@net.bio.net. Leave the Subject: line blank (anything entered on the Subject: line is ignored). 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Tue Nov 14 22:00:00 1995 Path: biosci!THALAMUS.WUSTL.EDU!navejoy From: navejoy@THALAMUS.WUSTL.EDU (Joy Nave Mentzer) Newsgroups: bionet.molbio.proteins.7tms_r Subject: mutant GPCR database Date: 15 Nov 1995 10:24:24 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511151826.MAA26731@thalamus.wustl.edu> NNTP-Posting-Host: net.bio.net I'm looking for a database (containing structural, functional &/or binding data) for mutant G protein coupled receptors. Does such an animal exist? Any assistance that could be provided is much appreciated. Thanks in advance, Joy Joy M. Nave Mentzer, Ph.D. Pfizer Postdoctoral Research Fellow Department of Anatomy and Neurobiology Washington University School of Medicine Campus Box 8108 660 S. Euclid Ave. St. Louis, MO 63110-1093 Phone: (314) 362-3468 FAX: (314) 362-3446 E-mail: navejoy@thalamus.wustl.edu From owner-7tms_r@net.bio.net Wed Nov 15 22:00:00 1995 Path: biosci!novo.dk!byw From: byw@novo.dk (Robert Bywater) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 16 Nov 1995 06:23:03 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 39 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511161418.PAA01691@bach> NNTP-Posting-Host: net.bio.net Hi 7TM netters Is there any good, recent review or informative paper on fluorescence studies in GPCRs of the opsin family ? Specifically for those GPCRs that do NOT have peptide ligands. ( I am aware of the elegant work of Andre Chollet, but that seems to be mostly concerned with NK2 receptors and the like. I was thinking more in terms of adrenergic/dopaminergic/serotonin subfamilies ... ). thanX for any help, and if I get a lot of interesting references I'll summarize. Robert Bywater ****************************** * * * Robert Bywater * * * * Biostructure Department * * NOVO NORDISK A/S * * * * DK-2880 BAGSVAERD Denmark * * * * email byw@novo.dk * * fax :: +45 4442 1400 * ****************************** From owner-7tms_r@net.bio.net Wed Nov 15 22:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!sdd.hp.com!swrinde!newsfeed.internetmci.com!in1.uu.net!csn!nntp-xfer-2.csn.net!yuma!purdue!news.bu.edu!acs4.bu.edu!albazie From: albazie@bu.edu (Saleh Al-Bazie) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Looking for anti-rabbit antibodies Date: 16 Nov 1995 16:14:40 GMT Organization: Boston University Lines: 10 Message-ID: <48fo1g$sav@news.bu.edu> NNTP-Posting-Host: acs4.bu.edu X-Newsreader: TIN [version 1.2 PL2] Subject: Looking for anti-rabbit antibodies Newsgroups: bionet.molbio.proteins.7tms_r Summary: Keywords: We are looking for 1) Anti-rabbit monoclonal antibodies against alkaline phosphatase and 2) Anti-rabbit monoclonal antibodies against osteocalcin. Thank you very much. From owner-7tms_r@net.bio.net Wed Nov 15 22:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!news.uoregon.edu!news.corpcomm.net!newstand.syr.edu!newsstand.cit.cornell.edu!NewsWatcher!user From: tpd4@cornell.edu (Terry Delaney) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: mutant GPCR database Date: Thu, 16 Nov 1995 09:38:54 -0500 Organization: Cornell University Lines: 31 Sender: tpd4@cornell.edu (Verified) Distribution: world Message-ID: References: <199511151826.MAA26731@thalamus.wustl.edu> NNTP-Posting-Host: 132 In article <199511151826.MAA26731@thalamus.wustl.edu>, navejoy@THALAMUS.WUSTL.EDU (Joy Nave Mentzer) wrote: > I'm looking for a database (containing structural, functional &/or binding > data) for mutant G protein coupled receptors. > clip > Joy M. Nave Mentzer, Ph.D. > Department of Anatomy and Neurobiology > Washington University School of Medicine Are you aware of Frank Kolakowski's G protein-Coupled Receptor DataBase website? He is at the Univ of Texas Health Science Center? The URL is: http://receptor.mgh.harvard.edu/GCRDBHOME.html. I would appreciate learning of other responses you receive to your query. Regards, Terry Delaney mmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmm Terrence P. Delaney Department of Plant Pathology Cornell University (607) 255-7856 334 Plant Science Building FAX (607) 255-4471 Ithaca, NY 14853-4203 tpd4@cornell.edu http://ppathw3.cals.cornell.edu/ppath/Delaney/Delaney.html mmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmmwmm From owner-7tms_r@net.bio.net Thu Nov 16 22:00:00 1995 Path: biosci!BIOCSERVER.BIOC.CWRU.EDU!roth From: roth@BIOCSERVER.BIOC.CWRU.EDU (Bryan Roth) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Joyce Baldwin/Henderson Date: 17 Nov 1995 11:24:27 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511171922.TAA03437@biocserver.BIOC.CWRU.Edu> NNTP-Posting-Host: net.bio.net I'm trying to get in contact with either Joyce Baldwin or Henderson. Does anyone out there know their e-mail addresses? Thanks! ============================================================= Bryan L.Roth Department of Biochemistry Room W438 CWRU Medical School 10900 Euclid Avenue Cleveland, OH 44106-4935 roth@biocserver.cwru.edu 216-368-2730 (Office) 216-368-4544 (FAX) From owner-7tms_r@net.bio.net Thu Nov 16 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bcm.tmc.edu!cs.utexas.edu!convex!darwin.sura.net!nih-csl!helix.nih.gov!dewolf From: "A.M. van Rhee" Subject: Re: mutant GPCR database In-Reply-To: Content-Type: TEXT/PLAIN; charset=US-ASCII Message-ID: Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: helix.nih.gov Organization: National Institutes of Health References: <199511151826.MAA26731@thalamus.wustl.edu> Mime-Version: 1.0 Date: Fri, 17 Nov 1995 15:01:25 GMT Lines: 43 Hi all, On Thu, 16 Nov 1995, Terry Delaney wrote: > In article <199511151826.MAA26731@thalamus.wustl.edu>, > navejoy@THALAMUS.WUSTL.EDU (Joy Nave Mentzer) wrote: > > > I'm looking for a database (containing structural, functional &/or binding > > data) for mutant G protein coupled receptors. > > > > clip > > > Joy M. Nave Mentzer, Ph.D. > > Department of Anatomy and Neurobiology > > Washington University School of Medicine here at NIH we have compiled an alignment of all known opsin related GPCRs, and we are in the process of compiling a mutation database. We're hoping to have the database up and running before year's end. For now you can retrieve any GPCR sequence through our website, and as soon as our paper with regard to the mutation database is accepted we'll try to get that part of our work on-line as well. Any depositions of mutations (preferably published) are greatly welcomed! Just thought I'd mention it. Sorry I can't help you right away. Regards, Michael. Dr. A.M. van Rhee National Institutes of Health National Institute of Diabetes, Digestive and Kidney Diseases Laboratory of Bioorganic Chemistry - Molecular Recognition Section Bldg. 8A Rm. B1A17 Bethesda, Maryland 20892-0810 tel: (301) 435-1936 // fax: (301) 402-4182 // dewolf@helix.nih.gov URL: http://mgddk1.niddk.nih.gov:8000/ From owner-7tms_r@net.bio.net Thu Nov 16 22:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!swrinde!tank.news.pipex.net!pipex!demon!sunsite.doc.ic.ac.uk!daresbury!not-for-mail From: Newsgroups: bionet.molbio.proteins.7tms_r Subject: Phosphorylation and its consequences. Date: 17 Nov 1995 14:53:23 -0000 Lines: 15 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <48i7l3$bku@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk I have some questions about phosphorylation of GPCRs at places other than the C terminus. 1. Is there evidence that GPCRs can or cannot be phosphorylated (other than the C-term)? If so, where? 2. Does phosphorylation ever occur at the spot in the beta-adrenergic receptor that Lefkowitz showed when mutated produced constitutively active GPCR at the beginnng of helix 6? 3. What are the functional consequences of phosphorylation? 4. Has a GPCR been shown to be a substrate for a specific kinase that phosphorylates at a site other than the C-terminus (that would leave out rhodopsin kinase)? Thanks very much. Martha Teeter, on sabbatical at the Max-Planck-Institut fuer Biophysik Frankfurt, Germany From owner-7tms_r@net.bio.net Mon Nov 20 22:00:00 1995 Path: biosci!agate!usenet.kornet.nm.kr!news.kreonet.re.kr!news.dacom.co.kr!simtel!torn!newshost.uwo.ca!usenet From: "Michael W. Clarke" Newsgroups: bionet.molbio.proteins.7tms_r Subject: (no subject) Date: 21 Nov 1995 14:51:13 GMT Organization: University of Western Ontario Lines: 12 Message-ID: <48sp11$mq9@falcon.ccs.uwo.ca> NNTP-Posting-Host: africa.mni.uwo.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1IS (X11; I; IRIX 5.3 IP22) X-URL: news:bionet.molbio.proteins.7tms_r Can anyone tell me if there is a site or a search strategy which will allow one to obtain all amino acid sequences of all known 7 transmembrane domain proteins? Thanks, Michael Clarke, PhD -- Department of Microbiology and Immunology University of Western Ontario London, ON N6A 5C1 Canada From owner-7tms_r@net.bio.net Mon Nov 20 22:00:00 1995 Path: biosci!VAXA.CRC.MSSM.EDU!strahs From: strahs@VAXA.CRC.MSSM.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: RE: (no subject) Date: 21 Nov 1995 09:23:33 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 34 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511211723.JAA14136@net.bio.net> NNTP-Posting-Host: net.bio.net >Can anyone tell me if there is a site or a search strategy which will allow one >to obtain all amino acid sequences of all known 7 transmembrane domain >proteins? > >Thanks, >Michael Clarke, PhD a few suggestions: your choice depends on how much time you want to spend and how much you really mean those words "all known"... 1) Run a BLAST or FASTA search with a 7tm receptor. Store all the responses. Pick one of the least homologous sequences retrieved. Fire that off on a new round of BLAST/FASTA searches. After a few rounds of this, you'll recover most of the sequence space of 7tm receptors. To speed up the process, start with a disjoint set of receptors (such as beta2-adrenergic, somatostatin, calcitonin and rhodopsin). You'll still miss things, but not much. 2) use the WWW site maintained by Lee Kolakowski. The address is http://receptor.mgh.harvard.edu/ . Again, this isn't absolutely complete, but... 3) Go to the EMBL server. They maintain some 7tm lists there. The address is http://www.sander.embl-heidelberg.de/7tm/ . ibid. 4) Use a scheme to retrieve the receptors you would like. An implementable scheme is described in Attwood TK. Findlay JB Fingerprinting G-protein-coupled receptors. Protein Engineering. 7(2):195-203, 1994 Feb. x From owner-7tms_r@net.bio.net Mon Nov 20 22:00:00 1995 Path: biosci!bath.ac.uk!bspma From: bspma@bath.ac.uk (M Aliste) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribed Date: 21 Nov 1995 11:00:59 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net subscribed From owner-7tms_r@net.bio.net Mon Nov 20 22:00:00 1995 Path: biosci!aecom.yu.edu!angelett From: angelett@aecom.yu.edu (Ruth Hogue Angeletti) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: (no subject) Date: 21 Nov 1995 12:13:10 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511211946.AA21243@alsys1.aecom.yu.edu> NNTP-Posting-Host: net.bio.net The sequences can be downloaded from an EMBL database in Heidelberg by anonymouse ftp, as described in the following reference: L Oliveira, ACM Paiva, C Sander, G Vriend (1994) A common step for signal transduction in G protein-coupled receptors. TIPS 15, 170-172. >Can anyone tell me if there is a site or a search strategy which will allow one >to obtain all amino acid sequences of all known 7 transmembrane domain >proteins? > >Thanks, >Michael Clarke, PhD >-- >Department of Microbiology and Immunology >University of Western Ontario >London, ON N6A 5C1 >Canada > > > Ruth Hogue Angeletti Albert Einstein College of Medicine 1300 Morris Park Avenue Bronx NY 10461 USA tel: 718-430-3475 fax: 718-430-8567 angelett@aecom.yu.edu From owner-7tms_r@net.bio.net Tue Nov 21 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!Germany.EU.net!news.dfn.de!news.ruhr-uni-bochum.de!news.rhrz.uni-bonn.de!news.uni-stuttgart.de!uni-regensburg.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Fluorescence studies on GPCRs Date: 21 Nov 1995 20:57:31 GMT Organization: Dept. of Pharmacology, U Wuerzburg Lines: 34 Message-ID: <48tefr$acl@winx03.informatik.uni-wuerzburg.de> References: <199511161418.PAA01691@bach> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Probably due to another failure of our news server :-( I have missed a posting by byw@novo.dk (Robert Bywater) where he asked about fluorescence studies on G protein-coupled receptors. I am not exactly sure what he wants to know. There have been experiments with fluorescent ligands, and in fact, I have just heard a talk of Prof. MacGrath from Glasgow who does confocal microscopy with both fluorescent prazosine and CGP-xxxxx derivates (insert correct number for xxxxx here). The preparation of the latter ligand has been described in @article{heithier:94, author = {Helmut Heithier and Dieter Hallmann and Fritz Boege and Helmut Reil"ander and Christian Dees and Knut A. Jaeggi and Donna Arndt-Jovin and Thomas M. Jovin and Ernst J. M. Helmreich}, title = {Synthesis and Properties of Fluorescent $\beta$-Adrenoceptor Ligands.}, journal = {Biochemistry}, volume = 33, pages = {9126--9134}, year = 1994 } Prof. MacGrath mentioned in his talk that Brian Kobilka had published recently a paper on modification of the recombinantly expressed beta2- adrenergic receptor itself with fluorescent groups. I am not aware of this publication but would appreciate any reference. --Cornelius (struggling with the university's news server). -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Tue Nov 21 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!Germany.EU.net!news.dfn.de!news.ruhr-uni-bochum.de!news.rhrz.uni-bonn.de!news.uni-stuttgart.de!uni-regensburg.de!lrz-muenchen.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Sucrose monolaureate Date: 21 Nov 1995 20:47:38 GMT Organization: Dept. of Pharmacology, U Wuerzburg Lines: 16 Message-ID: <48tdta$acl@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Apparently we have some problems with our news server. I just found out from the gopher at net.bio.net that Tuomo Glumoff asked about the usage of sucrose monolaureate in solubilization of membrane proteins. We use laurylsucrose (which is AFAIK the same) in the purification of recombinant beta2-adrenergic receptors expressed in Sf9 cells. The cell membranes are solubilized in laurylsucrose which is later exchanged with dodecylmaltoside. --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Tue Nov 21 22:00:00 1995 Path: biosci!ccmail.med.nyu.edu!ERIC.SIMON From: ERIC.SIMON@ccmail.med.nyu.edu Newsgroups: bionet.molbio.proteins.7tms_r Subject: cDNA library Date: 22 Nov 1995 12:24:06 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 6 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HXY5LTP2HS8WW80D@MCRCR6.MED.NYU.EDU> NNTP-Posting-Host: net.bio.net Does anyone know where we could obtain a good, randomly primed cDNA library prepared from bovine brain. We have used a commercial one (Clontech) and have a partial sequence (mu opioid receptor), but cannot seem to get the rest. We want to try a new library. Thanks, Eric J. Simon From owner-7tms_r@net.bio.net Tue Nov 21 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!Germany.EU.net!news.dfn.de!news.ruhr-uni-bochum.de!news.rhrz.uni-bonn.de!news.uni-stuttgart.de!uni-regensburg.de!lrz-muenchen.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Phosphorylation and its consequences. Date: 21 Nov 1995 21:09:13 GMT Organization: Dept. of Pharmacology, U Wuerzburg Lines: 82 Distribution: bionet Message-ID: <48tf5p$acl@winx03.informatik.uni-wuerzburg.de> References: <48i7l3$bku@mserv1.dl.ac.uk> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] teeter@max.mpibp.uni-frankfurt.de wrote: > I have some questions about phosphorylation of GPCRs at places other than the > C terminus. > 1. Is there evidence that GPCRs can or cannot be phosphorylated (other > than the C-term)? If so, where? There are several reports on phosphorylation of GPCRs in the third intracellular loop by receptor kinases (GRKs). PKA and PKC have also sites in the beta2-AR which might serve as substrates but these are apparently not used. I don't know anything about research on those phosphorylation sites in other receptors. In addition, there exists the possibility that GPCRs are phosphorylated by tyrosine kinases, but phosphorylation sites have so far not been mapped (AFAIK). > 2. Does phosphorylation ever occur at the spot in the beta-adrenergic > receptor that Lefkowitz showed when mutated produced constitutively active > GPCR at the beginnng of helix 6? AFAIK nothing in this way has ever been demonstrated. When receptor phosphorylation in the 3rd intracellular loop was shown, it usually occured roughly in the middle of the loop. > 3. What are the functional consequences of phosphorylation? Phosphorylation by PKA/PKC leads to direct uncoupling between receptor and G proteins, whereas phosphorylation by GRKs leads to binding of arrestin-like proteins which in turn impaires coupling between the receptor and the G protein. > 4. Has a GPCR been shown to be a substrate for a specific kinase that > phosphorylates at a site other than the C-terminus (that would leave out > rhodopsin kinase)? During a quick search of my literature folders I found the following papers: @article{nakata:94, author = {Hiroko Nakata and Kimihiko Kameyama and Kazuko Haga and Tatsuya Haga}, title = {Location of agonist-dependent phosphorylation sites in the third intracellular loop of muscarinic acetylcholine receptors (m2 subtypes).}, journal = {Eur. J. Biochem.}, volume = 220, pages = {29--36}, year = 1994 } @article{eason:95, author = {Margaret G. Eason and Sandra P. Moreira and Stephen B. Liggett}, title = {Four Consecutive Serines in the Third Intracellular Loop Are the Sites for $\beta$-Adrenergic Receptor Kinase-mediated Phosphorylation and Desensitization of the $\alpha_{{\rm 2A}}$-Adrenergic Receptor.}, journal = {J. Biol. Chem.}, volume = 270, number = 9, pages = {4681--4688}, year = 1995 } @article{jewellmotz:95, author = {Elizabeth A. Jewell-Motz and Stephen B. Liggett}, title = {An Acidic Motif within the Third Intracellular Loop of the $\alpha_2${C}2 Adrenergic Receptor Is Required for Agonist-Promoted Phosphorylation and Desensitization.}, journal = {Biochemistry}, volume = 34, pages = {11946--11953}, year = 1995 } Hope that helps, --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Sun Nov 26 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!psgrain!nntp.teleport.com!usenet From: Kim Neve Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Phosphorylation and its consequences. Date: Mon, 27 Nov 1995 13:37:19 -0800 Organization: Portland VA Medical Center Lines: 37 Message-ID: <30BA2F8F.76CD@teleport.com> References: <48i7l3$bku@mserv1.dl.ac.uk> NNTP-Posting-Host: ip-pdx04-21.teleport.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b2a (Windows; I; 16bit) teeter@max.mpibp.uni-frankfurt.de wrote: > > I have some questions about phosphorylation of GPCRs at places other than the > C terminus. > 1. Is there evidence that GPCRs can or cannot be phosphorylated (other > than the C-term)? If so, where? Yes, Bouvier et al. (JBC 264:16786-16792, 1989) showed that the human beta2-adrenergic receptor is phosphorylated by PKA at Ser-261, towards the C-terminal end of the third cytoplasmic loop. > 2. Does phosphorylation ever occur at the spot in the beta-adrenergic > receptor that Lefkowitz showed when mutated produced constitutively active > GPCR at the beginnng of helix 6? It turns out that Ser-261 is several residues prior to the mutated region of the constitutively active B2AR, which begins at around AA 266, I think. > 3. What are the functional consequences of phosphorylation? Involved in heterologous (cAMP-dependent) desensitization of the receptors. > 4. Has a GPCR been shown to be a substrate for a specific kinase that > phosphorylates at a site other than the C-terminus (that would leave out > rhodopsin kinase)? Yes, PKA (as mentioned above). Kim Neve Portland VA Medical Center From owner-7tms_r@net.bio.net Sun Nov 26 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!avalon.net!news From: c:\winapps\avalon\qvt Newsgroups: bionet.molbio.proteins.7tms_r,bionet.molbio.proteins.fluorescent,bionet.molbio.yeastbionet.molec- Subject: New WWW Site for Researchers Date: 27 Nov 1995 23:32:47 GMT Organization: Integrated DNA Technologies, Inc. Lines: 14 Distribution: world Message-ID: <49dhqv$388@arthur.avalon.net> NNTP-Posting-Host: 205.217.138.69 Keywords: oligo DNA synthesis IDT X-Newsreader: Xref: biosci bionet.molbio.proteins.7tms_r:514 bionet.molbio.proteins.fluorescent:151 Researchers: Please note that our WWW site is fully active, and that many of the researchers could benefit from its use. There is an online ordering system for Custom DNA Synthesis, Technical Bulletins of all types, links to almost all of the other Biotech related Sites on the internet, and an online catalog. Give us a try and see if there is anything you can use! http://www.idtdna.com A.R. Warner Integrated DNA Technologies, Inc. IDT Advantage From owner-7tms_r@net.bio.net Sun Nov 26 22:00:00 1995 Path: biosci!novo.dk!byw From: byw@novo.dk (Robert Bywater) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 27 Nov 1995 02:03:31 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 147 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511270958.KAA13667@bach> NNTP-Posting-Host: net.bio.net Dear 7TM netters Here is a summary of the replies that I received to the enquiry that I posted earlier : >Is there any good, recent review or informative paper on >fluorescence studies in GPCRs of the opsin family ? Specifically >for those GPCRs that do NOT have peptide ligands. ( I am aware >of the elegant work of Andre Chollet, but that seems to be mostly >concerned with NK2 receptors and the like. I was thinking more >in terms of adrenergic/dopaminergic/serotonin subfamilies ... ). Many thanX to : Stuart Sealfon, Rick Neubig, Eric Parker, Cornelius Krasel and to Frank Kolakowski/Dave Kristofferson for managing this net service. Robert Bywater -------------------------------------- -------------------------------------- Brian Kobilka's lab has done some wonderful adrenergic receptor fluorescence work which is in press in JBC. Stuart Sealfon -------------------------------------- -------------------------------------- I'm afraid that there isn't that much on fluorescence studies of GPCR's for non-peptide ligands other than epitope tagging and antibodies. We tried to develop fluorescent probes for alpha2 agonist binding but never were able to get good affinity and fluorescence in the same molecule. A couple papers you might look at are: Tota, M.R. and Strader, C.D. Characterization of the binding domain of the b-adrenergic receptor with the fluorescent antagonist carazolol. Evidence for a buried ligand binding site. J.Biol.Chem. 265:16891-16897, 1990. Sklar, L.A., Fay, S.P., Seligmann, B.E., Freer, R.J., Muthukumaraswamy, N., and Mueller, H. Fluorescence analysis of the size of a binding pocket of a peptide receptor at natural abundance. Biochemistry 29:313-316, 1990. Moench, S.J.; Terry, C.E.; Dewey, T.G. Fluorescence labeling of the palmitoylation sites of rhodopsin. Biochemistry 33:5783-5790; 1994. Brian Kobilka has started to directly label beta receptors and a postdoc presented some functional data at the recent alpha2 meeting. They saw agonist specific effects but the kinetics were rather slow. We have been focussing recently on labelling G proteins to use as a readout for receptor activation. Here are a few references from that work. Kwon, G.; Axelrod, D.; Neubig, R.R. Lateral mobility of tetramethylrhodamine-labelled G protein a and bt subunits in NG 108-15 cells. Cell. Signal. 6:663-679; 1994. Kwon, G.; Remmers, A.E.; Datta, S.; Neubig, R.R. Synthesis and characterization of fluorescently labelled subunits of brain G protein. Biochemistry 32:2401-2408; 1993. Neubig, R.R.; Connolly, M.C.; Remmers, A.E. Rapid kinetics of G protein subunit association: a rate-limiting conformational change? FEBS Lett. 355:251-253; 1994. Remmers, A.; Neubig, R.R. Resonance energy transfer from fluorescent G protein subunits to membrane lipids. Biochemistry 32:2409-2414; 1993. Remmers, A.E.; Neubig, R.R. Differential G protein activation by fluoresecnt guanine nucleotide derivatives. FASEB J. 9:A391 1995. (Abstract) Remmers, A.E.; Posner, R.; Neubig, R.R. Fluorescent guanine nucleotide analogs and G protein activation. J. Biol. Chem. 269:13771-13778; 1994. I'll be interested to hear what else you come up with. Rick _________________________________________________________ Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450 -------------------------------------- -------------------------------------- Brian Kobilka presented an abstract at the recent Society for Neurosciences meeting on such studies on adrenergic receptors. It isn't published yet, but you might want to contact him if you are interested. Eric -------------------------------------- -------------------------------------- There have been experiments with fluorescent ligands, and in fact, I have just heard a talk of Prof. MacGrath from Glasgow who does confocal microscopy with both fluorescent prazosine and CGP-xxxxx derivates (insert correct number for xxxxx here). The preparation of the latter ligand has been described in @article{heithier:94, author = {Helmut Heithier and Dieter Hallmann and Fritz Boege and Helmut Reil"ander and Christian Dees and Knut A. Jaeggi and Donna Arndt-Jovin and Thomas M. Jovin and Ernst J. M. Helmreich}, title = {Synthesis and Properties of Fluorescent $\beta$-Adrenoceptor Ligands.}, journal = {Biochemistry}, volume = 33, pages = {9126--9134}, year = 1994 } Prof. MacGrath mentioned in his talk that Brian Kobilka had published recently a paper on modification of the recombinantly expressed beta2- adrenergic receptor itself with fluorescent groups. I am not aware of this publication but would appreciate any reference. Cornelius Krasel -------------------------------------- -------------------------------------- ****************************** * * * Robert Bywater * * * * Biostructure Department * * NOVO NORDISK A/S * * * * DK-2880 BAGSVAERD Denmark * * * * email byw@novo.dk * * fax :: +45 4442 1400 * ****************************** From owner-7tms_r@net.bio.net Sun Nov 26 22:00:00 1995 Path: biosci!bath.ac.uk!bsscw From: bsscw@bath.ac.uk (C Wood) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 27 Nov 1995 05:08:38 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 2 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <48tefr$acl@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: net.bio.net subscribe From owner-7tms_r@net.bio.net Wed Nov 29 22:00:00 1995 Path: biosci!MOOSE.UVM.EDU!craper From: craper@MOOSE.UVM.EDU (Cardy Raper) Newsgroups: bionet.molbio.proteins.7tms_r Subject: postdoc position available Date: 30 Nov 1995 10:53:47 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 44 Sender: daemon@net.bio.net Distribution: world Message-ID: <199511301852.NAA138424@moose.uvm.edu> NNTP-Posting-Host: net.bio.net SEEKING POSTDOCTORAL ASSOCIATE IN FUNGAL MOLECULAR GENETICS to study self/nonself recognition in a filamentous fungus with multiple mating types. Funding available for at least three years. Start date could be as early as January '96. Qualified candidates must have experience in genetics and molecular biology. We have isolated and sequenced several pheromone and pheromone receptor genes that reside in the two multispecific B mating-type loci of the Basidiomycete Schizophyllum commune. These genes encode molecules for recognition of self versus many nonselves resulting in the initiation of a defined pathway of sexual development leading to dikaryosis and mushroom production. Many mutants are available to study this process at the molecular level. This research involves DNA sequencing and sequence analyses, PCR amplification and cloning, nucleic acid hybridizations, in vitro mutagenesis, DNA mediated transformation, and identification of genes thought to be part of a signal transduction pathway, downstream of the pheromone/receptor interactions. Experiments designed to identify and locate gene products in the cell are also planned. Our lab is located in a new building, fully supplied with the necessary and latest equipment to carry out this research. The Department, which has tripled in size over the past several years, includes 24 faculty members representing a wide variety of interests in molecular genetics including signal transduction, morphogenesis and cell-cycle control in yeast, mechanisms of DNA damage and repair, transcription complexes, and ribozyme function. It provides a cooperative learning environment for students and postdocs at all levels. If interested, please send letter of application, curriculum vitae, names and addresses -- including e-mail address and/or phone numbers -- of three persons who can evaluate you as a candidate. If preferred, reply using e-mail address. Dr. Carlene A. Raper (craper@moose.uvm.edu) Dept. Microbiology and Molecular Genetics The L. P. Markey Center for Molecular Genetics Stafford Hall, Tel. (802) 656-1115; Fax. (802) 656-8749; University of Vermont, Burlington VT 05405 For published information, look for Wendland et al, 1995, EMBO Journal, vol 14, no. 21, pp. 5271-5278 From owner-7tms_r@net.bio.net Thu Nov 30 22:00:00 1995 Path: biosci!UCONNVM.UCONN.EDU!GMS95003 From: GMS95003@UCONNVM.UCONN.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 1 Dec 1995 11:03:01 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <951201.140108.EST.GMS95003@UConnVM.UConn.Edu> NNTP-Posting-Host: net.bio.net subscribe 7tms_r From owner-7tms_r@net.bio.net Thu Nov 30 22:00:00 1995 Path: biosci!sci.himeji-tech.ac.jp!nakagawa From: nakagawa@sci.himeji-tech.ac.jp (Masashi NAKAGAWA) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 1 Dec 1995 15:56:45 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <199512020000.JAA22788@perm.sci.himeji-tech.ac.jp> NNTP-Posting-Host: net.bio.net help ******************************** *　　Masashi NAKAGAWA　　　　　　　　　　　* *Department of Life Science, Faculty of Scienc * *　Himeji Institute of Technology 　　 *　 *　@e-mail: nakagawa@sci.himeji-tech.ac.jp * * Tel: 07915-8-0195 * * Fax: 07915-8-0197 * ********************************